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HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY
From the Center for Molecular and Vascular Biology,
University of Leuven, Belgium; and Academic Department of Surgery, St
Thomas' Hospital, London, United Kingdom.
The human monoclonal antibody mAb-LE2E9 partially inactivates human
factor VIII (FVIII), leaving approximately 10% residual activity. The
antithrombotic efficacy of the antibody was evaluated in mouse models
of inferior vena cava thrombosis. Thrombi were induced in wild-type
mice given either the antibody or saline. No thrombi occurred in any of
8 mice treated with mAb-LE2E9, whereas 6 of 8 control mice developed
thrombi (P = .007). Treatment with mAb-LE2E9 did not
result in a severe bleeding phenotype: a tail-cutting experiment that
resulted in death of C57BL/6 FVIII-deficient (FVIII Venous thromboembolic disease remains a major
health issue, with an incidence of 1 to 3 per 1000 individuals per year
and a high early mortality rate.1,2 Current antithrombotic
therapies primarily consist of heparin (or low molecular weight
heparins) and vitamin K antagonists. All carry a significant risk of
bleeding,3 which limits both the dose and duration of
treatment and may require regular monitoring.4,5 Safer and
more efficient methods for the prevention and treatment of venous
thromboembolic diseases are therefore desirable.
Factor VIII (FVIII), an essential cofactor of blood coagulation, is a
330-kd glycoprotein produced by the liver, which circulates in the form
of a complex with von Willebrand factor (VWF). Upon cleavage by
thrombin, activated FVIII (FVIIIa) dissociates from VWF and assembles
with activated factor IX (FIXa) on negatively charged phospholipids to
form the "tenase" complex, resulting in a 100 000-fold increased
activation of factor X.6 Besides its key role in
hemostasis, FVIII has been associated with the development of venous
thrombosis,7-10 suggesting that FVIII could constitute an
appropriate target for antithrombotic therapy.
The human high-affinity monoclonal antibody mAb-LE2E9 inhibits the
procoagulant activity of FVIII specifically, but only partially (around
90%), even in large molar excess.11 Here, we hypothesized that administration of mAb-LE2E9 might have an antithrombotic effect
without the risk of overdosing or spontaneous hemorrhage. The aim of
this study was to evaluate the pharmacokinetics and potential
antithrombotic effect of mAb-LE2E9 in a novel model of venous thrombosis.
Reagents
FVIII assays
In vitro inhibition of rhFVIII was also measured when various concentrations of rhFVIII were incubated with mAb-LE2E9 at a fixed concentration and in excess over FVIII. Thus, rhFVIII at final concentrations of 1, 2, 4, 8, and 16 IU/mL was added to plasma preincubated for 2 hours at 37°C with 1 or 20 µg/mL mAb-LE2E9 or control buffer. Following a further 2-hour incubation period at 37°C, residual FVIII activity was measured in a chromogenic substrate assay. Pharmacokinetics of mAb-LE2E9 in wild-type mice The plasma disappearance rate of 100 µg intravenously or subcutaneously injected mAb-LE2E9 (which constitutes at least 100-fold molar excess over murine FVIII) and its effect on circulating murine FVIII levels were measured in blood samples obtained by cardiac puncture in groups of 3 mice taken at intervals up to 3 weeks after injection. Levels of mAb-LE2E9 were measured by enzyme-linked immunosorbent assay and expressed in percentage of the value obtained 20 minutes after intravenous injection and 24 hours after subcutaneous administration.Pharmacokinetics of rhFVIII, mAb-LE2E9, and mAb-LE2E9/rhFVIII
complex in FVIII  / mice were determined following
intravenous injection of (1) 7.5 IU rhFVIII alone, (2) 7.5 IU rhFVIII
complexed with 10 µg mAb-LE2E9, and (3) 7.5 IU rhFVIII followed 15 minutes later by 10 µg mAb-LE2E9.
Bolus tail vein injections of 100 µL were used. Levels of rhFVIII were measured in blood samples collected by cardiac puncture at 5, 10, 15, 30, 45, 260, and 420 minutes after injection (n = 3 mice per time point). Mouse model of vena cava thrombosis in wild-type mice Thrombus was produced in the inferior vena cava of adult male wild-type mice (weight, 18-31 g; age, 8-10 weeks) using a modification of the previously described model of vena cava thrombosis in the rat.13 Mice were anesthetized with isoflurane, the inferior vena cava was exposed below the renal veins via a median laparotomy, and a neurosurgical vascular clip (Braun Medical, Sheffield, England) was applied for 15 seconds on 2 occasions, 30 seconds apart, to a segment of the vena cava. A 5/0 Prolene thread was then placed alongside the vena cava and a stenosis produced by tying a 4/0 silk suture around the vena cava and the Prolene thread. The thread was removed to allow blood flow to resume. The abdomen was closed and the animal allowed to recover. After 4 hours, the mice were reanesthetized and a 1 cm portion of the inferior vena cava (between the point of ligature and iliac bifurcation) was excised, washed in 10% phosphate-buffered saline, and soaked overnight in 1% paraformaldehyde. Vessel segments were embedded in paraffin wax, and 7-µm transverse sections were cut at 0.5-mm intervals from the ligature down.Thrombus assessment Sections were stained by hematoxylin and eosin, Martius Scarlet Blue (MSB), and a rabbit antiplatelet antibody (Accurate Chemical & Scientific, Westbury, NY). MSB stains fresh fibrin red or mature fibrin blue/gray, red cells yellow, and collagen bright blue.Thrombus size was measured by scoring the 7 sections for the presence of thrombus, giving a score of 1 for the presence and 0 for the absence of thrombus in each. Scores were then added up for each animal. The investigators performing the operations and the microscopic analyses were blinded toward treatment groups. Thrombus formation in wild-type mice treated with mAb-LE2E9 Thrombosis was induced in 2 groups of 8 wild-type mice 16 hours following subcutaneous injection of 150 µg mAb-LE2E9 or saline. In additional experiments, 2 groups of 6 wild-type C57BL/6 mice were treated with an intravenous injection of 10 µg mAb-LE2E9 or saline 30 minutes before thrombus induction.Effect of mAb-LE2E9 antibody on thrombus formation in
FVIII In the first experiments, venous thrombosis was induced in a group of
18 FVIII In the second set of experiments, a cohort of 28 FVIII All animals were killed after 4 hours, and 1 cm of the vena cava, between the point of ligature and iliac bifurcation, was harvested and processed for histology. Effect of mAb-LE2E9 on bleeding after tail cutting in wild-type mice The risk of severe bleeding associated with high concentrations of mAb-LE2E9 in plasma was evaluated in a tail-cutting experiment. This assay is based on the observation that sectioning of the distal portion of the tail results in important blood loss leading to death in most FVIII/ mice14 whereas normal mice
should survive. This procedure allows an in vivo evaluation of FVIII
activity. Twelve wild-type mice were injected subcutaneously with
either 150 µg mAb-LE2E9 (n = 6) or saline (n = 6). Tail cutting
was also performed in 5 FVIII/ mice in a control
experiment. A 7-mm section of the tail was then cut 16 hours later and
survival rate monitored over the subsequent 24 hours.
Statistical analysis The statistical significance of differences between groups was evaluated on the presence or absence of thrombus using the Fisher exact test (2-sided). The effect on thrombus size was tested by comparing thrombus scores using the Mann-Whitney U test.15
In vitro inhibition of FVIII by mAb-LE2E9 in plasma from different species To identify animal models suitable to evaluate the antithrombotic activity of mAb-LE2E9, inhibition of FVIII was measured in plasma of different strains. Rat and rabbit FVIII appeared to be resistant to inhibition by mAb-LE2E9. By contrast, mouse FVIII activity was dose-dependently inhibited, although with a lower efficacy, evidenced by a shift to the right of FVIII inhibition curve. Twenty times more mAb-LE2E9 was required to achieve maximal FVIII inhibition for murine than for human plasma.In vitro inhibition of rhFVIII was evaluated by incubating different concentrations of rhFVIII spiked in human plasma containing mAb-LE2E9. At rhFVIII concentrations between 1 and 8 IU/mL, the proportion of FVIII activity inhibited by 20 µg/mL mAb-LE2E9 was identical: the residual FVIII activity increased by 0.11 ± 0.03 IU/mL (mean ± SD) per international unit of rhFVIII added to plasma. Similarly, in presence of 1 µg/mL mAb-LE2E9, the increase in FVIII activity was 0.12 ± 0.02 IU/mL (mean ± SD) per international unit of rhFVIII added to plasma, when rhFVIII was added at concentrations lower or equal to 4 IU/mL. Pharmocokinetics and pharmacodynamics of mAb-LE2E9 in wild-type mice Because mouse FVIII was inhibited by mAb-LE2E9, this animal strain was selected for in vivo analysis of mAb-LE2E9 activity. Injection of 100 µg mAb-LE2E9 in wild-type mice, which constitutes at least 100-fold molar excess over murine FVIII, was followed by a plasma disappearance rate with a half-life of about 3 days. The murine endogenous FVIII decreased to roughly 10% of baseline for 7 days when the mAb-LE2E9 level had decreased by two thirds, after which FVIII gradually rose to reach baseline levels at 20 days, concomitant with further clearance of the antibody. No significant differences were observed between intravenous (Figure 1A) and subcutaneous (Figure 1B) injection of the antibody.
Characteristics of mouse model of venous thrombosis A new model was developed to evaluate the development of deep vein thrombosis in mice. Vascular wall damages and partial stenosis of the vena cava were induced by short time compression and ligation of the vein. Experimental thrombi that occluded up to 100% of the vessel were produced by 4 hours in 70% to 80% of wild-type mice and in FVIII/ mice reconstituted with rhFVIII. The procedure
was well tolerated (no evidence of hemorrhage and no postoperative
deaths). Thrombi had a classical coralline structure with red cells
trapped between layers of platelets, leukocytes, and fibrin, indicating
that they were formed under flow conditions (Figure
2A-C). This structure resembles that of
human venous thrombi.16
Effect of mAb-LE2E9 on thrombosis in wild-type mice Two groups of 8 mice were injected subcutaneously with either 150 µg mAb-LE2E9 or saline 16 hours before induction of thrombosis. Six of 8 mice injected with saline developed a thrombus, compared with 0 of 8 animals pretreated with mAb-LE2E9 (P = .007, Figure 3). By contrast, no significant reduction of thrombus was seen following intravenous administration of 10 µg mAb-LE2E9 into wild-type mice 30 minutes before induction of thrombosis (4 of 6 mice injected with saline and 3 of 6 mice treated with mAb-LE2E9 developed a thrombus).
Vena cava thrombosis in FVIII / mice reconstituted
with rhFVIII. We first determined whether induction of deep vein
thrombosis in FVIII/ mice required administration of
FVIII. The surgical procedure for induction of vena cava thrombus was
applied to 6 FVIII/ mice. All animals showed signs of
significant blood loss and deterioration of their physical condition by
4 hours after surgery, and none produced a thrombus. Therefore,
FVIII/ mice reconstituted with rhFVIII were explored as
an alternative model to their wild-type counterpart.
Pharmacokinetics of rhFVIII and rhFVIII/mAb-LE2E9 complex in
FVIII / mice were determined for rhFVIII, for rhFVIII
complexed in vitro with mAb-LE2E9, and for rhFVIII followed by
injection of mAb-LE2E9, as described in "Materials and methods."
Injection of 7.5 IU rhFVIII produced FVIII levels of 4.6 ± 1.2 IU/mL
(mean ± SD) at 30 minutes, which was cleared with a plasma half-life
of approximately 3 hours to reach levels of 1.3 ± 0.8 IU/mL after
4.5 hours (Figure 4A). Injection of 100 µL of a mixture of 7.5 IU rhFVIII, preincubated for 30 minutes with
10 µg mAb-LE2E9 (resulting in plateau inhibition), produced FVIII
levels of 0.32 ± 0.02 IU/mL at 30 minutes, which was cleared with a
plasma half-life of approximately 4 hours (Figure 4B). Finally,
injection of 7.5 rhFVIII followed 15 minutes later by injection of 10 µg mAb-LE2E9 produced FVIII levels at 15 minutes of 0.36 ± 0.04
IU/mL, which was cleared with a plasma half-life of approximately 4 hours (Figure 4C).
Vena cava thrombosis in FVIII / mice
following administration of 7.5 IU rhFVIII, which resulted in a
concentration of FVIII higher than 1 IU/mL for more than 4 hours.
Thrombi formed in FVIII/ mice reconstituted with
rhFVIII were microscopically similar to those formed in wild-type
animals (Figure 2D-F and 2A-C, respectively).
Effect of mAb-LE2E9 on thrombosis in FVIII
Recombinant human FVIIII preincubated with mAb-LE2E9. Thrombus was observed in 7 of 9 mice treated with rhFVIII only, compared with 1 of 9 mice in the rhFVIII/mAb-LE2E9 complex-treated group (P = .015) (Figure 5A). The median thrombus score in the rhFVIII group was 4 (interquartile range 1 to 7), compared with 0 (interquartile range 0 to 0) in the rhFVIII/mAb-LE2E9 group (P < .001). Recombinant human FVIIII followed by mAb-LE2E9. Thrombus was found in 10 of 14 mice in the rhFVIII-treated group but in only 1 of 14 mice treated with the rhFVIII followed by mAbLE2E9 (P < .001) (Figure 5B). The median thrombus scores were 6 (interquartile range 0 to 7) and 0 (interquartile range 0 to 0), respectively (P < .01). Effect of mAb-LE2E9 on bleeding after tail cutting in wild-type mice The risk of severe bleeding associated with high concentrations of mAb-LE2E9 in plasma was evaluated in a tail-cutting experiment. This assay is based on the observation that sectioning of the distal portion of the tail results in important blood loss leading to death in most FVIII/ mice14 whereas normal mice survive.
This procedure therefore allows an in vivo evaluation of FVIII
activity. Thus, 6 mice were injected subcutaneously with 150 µg
mAb-LE2E9 16 hours before tail cutting. No death was recorded in
control wild-type mice or wild-type mice treated with mAb-LE2E9. In a
control group made of 5 FVIII/ mice, all animals died
within 24 hours after tail cutting.
In this study, we sought evidence that the partially inhibitory human monoclonal antibody against FVIII, mAb-LE2E9, might constitute a novel approach to anticoagulant therapy without the risk of overdosing or causing spontaneous bleeding. The possibility of using mAb-LE2E9 as an antithrombotic was derived from 2 main observations. (1) Patients with mild hemophilia A (FVIII levels between 5% and 40%) do not experience bleeding except after severe trauma or surgery.17 (2) Venous thrombosis is very rare in these patients.18 This observation supports a key role for FVIII in venous thrombosis.7-10 Partial inhibition of FVIII by mAb-LE2E9 seems to mimic mild hemophilia A. Reducing FVIII activity may limit coagulation while avoiding the risk of spontaneous hemorrhage. Some patients with hemophilia A develop an immune response toward FVIII
following administration of FVIII concentrates. The anti-FVIII
antibodies produced typically only partially inhibit FVIII activity in
presence of VWF, whereas in the absence of the latter, the
inhibition of FVIII is complete.19 By contrast, mAb-LE2E9 inhibits FVIII activity only partially even in
absence of VWF.11 The mechanism responsible for the
limited FVIII inactivation is still currently investigated. Surface
plasmon resonance analysis indicated that mAb-LE2E9 has a high affinity
for FVIII (kass = 2.9 × 105
M The evaluation of mAb-LE2E9 as an antithrombotic in existing
experimental rat and rabbit animal models has been hampered by the
species specificity of the human monoclonal antibody. Both rat and
rabbit FVIII are resistant to mAb-LE2E9. Mouse FVIII reacts, although
with lower affinity than human FVIII. The latter species has the
advantage that FVIII The experimental thrombi produced in this model consisted mainly of red
cells and were typically laminated with platelets, fibrin, and
leukocytes and were similar to those seen in the rat and in
humans.13,15 The thrombosis model also replicates several features relevant to the development of deep vein thrombosis in man,
including endothelial disturbance, low flow, and hypercoagulability (high levels of FVIII).7-10 Thrombus developed in about
70% to 80% of wild-type and FVIII Experiments were conducted both in wild-type mice and in
FVIII The results from this study show that in FVIII The surgical procedure did not cause excessive bleeding in animals
treated with mAb-LE2E9, while the FVIII The data in this study support the concept that anticoagulant agents targeting the intrinsic pathway of the coagulation cascade could be potent antithrombotic agents while maintaining a sufficient residual coagulation activity to preserve hemostasis. Several new anticoagulant agents targeting FIX have recently been developed.21,22 Anti-FIX antibodies also proved to be efficient in animal models of thrombosis while inducing only minor bleeding.23,24 However, mAb-LE2E9 has several advantages over these and other antithrombotic therapies. FVIII has the lowest plasma concentration of all the coagulation factors. A small amount of mAb-LE2E9 antibody will therefore be required for its inhibition. Monoclonal antibodies are currently used for treatment of chronic diseases, such as allergy.25,26 In the latter studies, the humanized antibody omalizumab was administered every other week for months at doses up to 300 mg per injection. Given the kinetics of FVIII inactivation by mAb-LE2E9, at least 10-fold less antibody would be required for patients with thrombotic disorders. IgG4 antibodies, such as mAb-LE2E9, have a half-life of 3 weeks in humans,27 which would reduce the frequency of administration and provide very stable plasma FVIII levels. Strong FVIII inhibition can be obtained while avoiding overdosing because, even in large excess, FVIII inhibition is only partial. Finally, administration of FVIII could be used to restore a normal coagulation level if required in case of trauma or surgery. In conclusion, our data show that a human monoclonal antibody that partially inhibits FVIII activity may represent a novel type of potent antithrombotic agent. This observation may have important implications for the development of efficient, easy, and safe strategies for the prevention and treatment of venous thrombosis.
Submitted September 5, 2001; accepted December 18, 2001.
Supported in part by grant G.0378.01 from the Flemish Research Foundation.
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Reprints: Marc Jacquemin, Center for Molecular and Vascular Biology, University of Leuven, Campus Gasthuisberg, Herestraat 49, B-3000 Leuven, Belgium; e-mail: marc.jacquemin{at}med.kuleuven.ac.be.
1. Nordstrom M, Lindblad B, Bergqvist D, Kjellstrom T. A prospective study of the incidence of deep-vein thrombosis within a defined urban population. J Intern Med. 1992;232:155-160[Medline] [Order article via Infotrieve]. 2. Rosendaal FR. Thrombosis in the young: epidemiology and risk factors. A focus on venous thrombosis. Thromb Haemost. 1997;78:1-6[Medline] [Order article via Infotrieve]. 3. Research Committee of the British Thoracic Society. Optimum duration of anticoagulation for deep-vein thrombosis and pulmonary embolism. Lancet. 1992;340:873-876[Medline] [Order article via Infotrieve].
4.
Hylek EM, Singer DE.
Risk factors for intracranial hemorrhage in outpatients taking warfarin.
Ann Intern Med.
1994;120:897-902
5.
Cannegieter SC, Rosendaal FR, Wintzen AR, van der Meer FJ, Vandenbroucke JP, Briet E.
Optimal oral anticoagulant therapy in patients with mechanical heart valves.
N Engl J Med.
1995;333:11-17
6.
van Dieijen G, Tans G, Rosing J, Hemker HC.
The role of phospholipid and factor VIIIa in the activation of bovine factor X.
J Biol Chem.
1981;256:3433-3442 7. Koster T, Blann AD, Briet E, Vandenbroucke JP, Rosendaal FR. Role of clotting factor VIII in effect of von Willebrand factor on occurrence of deep-vein thrombosis. Lancet. 1995;345:152-155[CrossRef][Medline] [Order article via Infotrieve].
8.
Kyrle PA, Minar E, Hirschl M, et al.
High plasma levels of factor VIII and the risk of recurrent venous thromboembolism.
N Engl J Med.
2000;343:457-462 9. O'Donnell J, Mumford AD, Manning RA, Laffan M. Elevation of FVIII: C in venous thromboembolism is persistent and independent of the acute phase response. Thromb Haemost. 2000;83:10-13[Medline] [Order article via Infotrieve]. 10. Kraaijenhagen RA, in't Anker PS, Koopman MM, et al. High plasma concentration of factor VIIIc is a major risk factor for venous thromboembolism. Thromb Haemost. 2000;83:5-9[Medline] [Order article via Infotrieve].
11.
Jacquemin M, Benhida A, Peerlinck K, et al.
A human antibody directed to the factor VIII C1 domain inhibits factor VIII cofactor activity and binding to von Willebrand factor.
Blood.
2000;95:156-163 12. Kasper CK, Aledort L, Aronson D, et al. Proceedings: a more uniform measurement of factor VIII inhibitors. Thromb Diath Haemorrh. 1975;34:869-872[Medline] [Order article via Infotrieve]. 13. Humphries J, Catharine L, McGuiness FR, et al. Monocyte chemotactic protein-1 (MCP-1) accelerates the organization and resolution of venous thrombi. J Vasc Surg. 1999;30:894-900[CrossRef][Medline] [Order article via Infotrieve]. 14. Bi L, Lawler AM, Antonarakis SE, High KA, Gearhart JD, Kazazian HH Jr. Targeted disruption of the mouse factor VIII gene produces a model of hemophilia A. Nat Genet. 1995;10:119-121[CrossRef][Medline] [Order article via Infotrieve]. 15. Blalock HM. Social Statistics. Rev 2nd ed. Tokyo, Japan: Mc-Graw-Hill Kogakusha; 1979. 16. Quarmby J, Smith A, Collins M, Cederholm-Williams S, Burnand K. A model of in vivo human venous thrombosis that confirms changes in the release of specific soluble cell adhesion molecules in experimental venous thrombogenesis. J Vasc Surg. 1999;30:139-147[CrossRef][Medline] [Order article via Infotrieve]. 17. Jones PK, Ratnoff OD. The changing prognosis of classic hemophilia (factor VIII "deficiency"). Ann Intern Med. 1991;114:641-648. 18. Stewart AJ, Manson LM, Dennis R, Allan PL, Ludlam CA. Thrombosis in a duplicated superficial femoral vein in a patient with hemophilia A. Hemophilia. 2000;6:47-49.
19.
Gawryl MS, Hoyer LW.
Inactivation of factor VIII coagulant activity by two different types of human antibodies.
Blood.
1982;60:1103-1109 20. Liu M, Thompson AR. Factor VIII's C1 domain enhances C2 binding of factors IX/IXa, X/Xa and von Willebrand factor [abstract]. Blood. 2000;96:489a.
21.
Choudhri TF, Hoh BL, Prestigiacomo CJ, et al.
Targeted inhibition of intrinsic coagulation limits cerebral injury in stroke without increasing intracerebral hemorrhage.
J Exp Med.
1999;190:91-99 22. Benedict CR, Ryan J, Wolitzky B, et al. Active site-blocked factor IXa prevents intravascular thrombus formation in the coronary vasculature without inhibiting extravascular coagulation in a canine thrombosis model. J Clin Invest. 1991;88:1760-1765.
23.
Refino CJ, Himber J, Burcklen L, et al.
A human antibody that binds to the
24.
Feuerstein GZ, Patel A, Toomey JR, et al.
Antithrombotic efficacy of a novel murine antihuman factor IX antibody in rats.
Arterioscler Thromb Vasc Biol.
1999;19:2554-2562 25. Milgrom H, Fick RB, Su JQ, et al. Treatment of allergic asthma with monoclonal anti-IgE antibody rhuMAb-E25 study group. N Engl J Med. 1999;26:1966-1973. 26. Boushey HA. Experiences with monoclonal antibody therapy for allergic asthma. J Allergy Clin Immunol. 2001;108:S77-S83[CrossRef][Medline] [Order article via Infotrieve]. 27. Morell A, Terry WD, Waldmann TA. Metabolic properties of IgG subclasses in man. J Clin Invest. 1970;49:673-680.
© 2002 by The American Society of Hematology.
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  D. Bonderman, J. Jakowitsch, B. Redwan, H. Bergmeister, M.-K. Renner, H. Panzenbock, C. Adlbrecht, A. Georgopoulos, W. Klepetko, M. Kneussl, et al. Role for Staphylococci in Misguided Thrombus Resolution of Chronic Thromboembolic Pulmonary Hypertension Arterioscler Thromb Vasc Biol, April 1, 2008; 28(4): 678 - 684. [Abstract] [Full Text] [PDF]
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 J. G. G. Gilles, S. C. Grailly, M. De Maeyer, M. G. Jacquemin, L. P. VanderElst, and J.-M. R. Saint-Remy In vivo neutralization of a C2 domain-specific human anti-Factor VIII inhibitor by an anti-idiotypic antibody Blood, April 1, 2004; 103(7): 2617 - 2623. [Abstract] [Full Text] [PDF]
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S. Villard, S. Lacroix-Desmazes, T. Kieber-Emmons, D. Piquer, S. Grailly, A. Benhida, S. V. Kaveri, J.-M. Saint-Remy, and C. Granier Peptide decoys selected by phage display block in vitro and in vivo activity of a human anti-FVIII inhibitor Blood, August 1, 2003; 102(3): 949 - 952. [Abstract] [Full Text] [PDF]
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 I. Singh, K.G. Burnand, M. Collins;, A. Luttun, D. Collen, B. Boelhouwer;, and A. Smith Failure of Thrombus to Resolve in Urokinase-Type Plasminogen Activator Gene-Knockout Mice: Rescue by Normal Bone Marrow-Derived Cells Circulation, February 18, 2003; 107(6): 869 - 875. [Abstract] [Full Text] [PDF]
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 S. Villard, D. Piquer, S. Raut, J.-P. Leonetti, J.-M. Saint-Remy, and C. Granier Low Molecular Weight Peptides Restore the Procoagulant Activity of Factor VIII in the Presence of the Potent Inhibitor Antibody ESH8 J. Biol. Chem., July 19, 2002; 277(30): 27232 - 27239. [Abstract] [Full Text] [PDF]
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Top
Abstract
Introduction
-carboxyglutamic acid domain of factor IX is a potent antithrombotic in vivo.
Thromb Haemost.
1999;82:1188-1195